Prebiotics-Controlled Disposable Engineered Bacteria for Intestinal Diseases

Abstract

As a new method of diagnosis and treatment for intestinal diseases, intelligent engineered bacteria based on synthetic biology have been developed vigorously in recent years. However, how to deal with the engineered bacteria in vivo after completing the tasks is an urgent problem to be resolved. In this study, we constructed a thiosulfate (a biomarker of inflammatory bowel disease)-responsive engineered bacteria to generate two signals, sfGFP (monitoring) and gain-of-function (translation activation) mutation (ACG to ATG), in the initiation codon of lysisE (recording) via the CRISPR/Cas9-mediated base editing system. Once these two signals were detected, xylose could be added to induce lysis E expression, resulting in the destruction of the edited bacteria and the release of AvCystain simultaneously. Overall, our innovative engineered bacteria can record instant and historical information of the disease, and especially, the edited bacteria can be artificially attenuated and release drug in situ when needed, ultimately serving as a disposable and recyclable candidate for more types of diseases.

Protocol for engineering E. coli Nissle 1917 to diagnose, record, and ameliorate inflammatory bowel disease in mice

“…However, we did not test this genetic circuit in other bacteria, except E. coli DH5α. Although both EcN 1 and DH5α 10 belong to Escherichia coli , their genetic circuits exhibit different performances. Therefore, we propose to consider some key points when applying thiosulfate-responsive sensors to other bacterial strains: 1.…”